Anti-Human Ki-67 (Ki-67) Colorwheel^® Dye-Ready mAb

ColorWheel^® technology is a novel and proprietary method of creating your own antibody and dye combinations for use in flow cytometry. By incubating any ColorWheel^® antibody with any ColorWheel^® dye, researchers can quickly and simply produce primary conjugated antibodies for use in single color or multicolor flow cytometry analysis. Each ColorWheel^® antibody and dye is lyophilized for long-term storage, allowing for simplicity and flexibility without compromise. ColorWheel^® technology requires both ColorWheel^® antibody and ColorWheel^® dye for Flow Cytometry application.

Proliferation marker protein Ki-67 (UniProt: P46013; also known as Antigen identified by monoclonal antibody Ki-67, KI-67, Ki67) is encoded by the MKI67 gene (Gene ID: 4288) in human. Ki67 antigen is the prototypic cell cycle related nuclear protein, expressed by proliferating cells in all phases of the active cell cycle (G1, S, G2 and M phase). In the G1 phase it is predominantly localized in the perinucleolar region and in the later phases it is also detected throughout the nuclear interior, being predominantly localized in the nuclear matrix. Its levels are shown to be the highest in the G2 phase and during mitosis. Ki67 is routinely used as a marker of cell cycling and proliferation. It is required to maintain individual mitotic chromosomes dispersed in the cytoplasm following nuclear envelope disassembly. It is shown to associate with the surface of the mitotic chromosome, the perichromosomal layer, and covers a substantial fraction of the chromosome surface and prevents chromosomes from collapsing into a single chromatin mass by forming a steric and electrostatic charge barrier. It binds DNA, with a preference for supercoiled DNA and AT-rich DNA. Ki67 undergoes phosphorylation and is shown to be hyperphosphorylated in mitosis phase. The hyperphosphorylated form fails to bind DNA. Major regions of the Ki-67 include an N-terminal forkhead-associated (FHA) domain (aa 27-76), a protein phosphatase 1 (PP1) binding domain (aa 502-549), a large central region comprising 16 tandem repeats, and a C-terminal leucine/arginine-rich chromatin-binding (LR) domain. Ki67 interacts with kinesin-like motor protein Kif15 and a nucleolar protein NIFK (Nucleolar Protein Interacting with the FHA domain of MKI67) via its FHA domain to promote cell proliferation and tumor metastasis. The central region with tandem repeats contains residues that are phosphorylated by CDK1 during mitosis. (Ref.: Sun, X., and Kaufman PD. (2018). Chromosoma. 127(2); 175-186).

We are committed to bringing you greener alternative products, which adhere to one or more of The 12 Principles of Green Chemistry.This product is Preservative-free, lyophilized product for enhanced stability and allow for ambient shipping and thus aligns with "Waste Prevention", "Designing Safer Chemicals" and "Design for Energy Efficiency". Click here for more information.

Nuclei of the Hodgkin lymphoma cell line L428.

Quality Control Testing

Evaluated by Flow Cytometry in Jurkat cells fixed and permeabilized with ice cold 70% ethanol.

Flow Cytometry Analysis (FC): Staining of one million fixed and permeabilized Jurkat cells was performed using 5 μL of a 1:1 mixture of Cat. No. CWA-1078, Anti-Human Ki-67 (Ki-67) ColorWheel^® Dye-Ready mAb and Cat. No. CWD-PE ColorWheel^® Antibody-Ready Phycoerythrin (PE) Dye or an equivalent amount of PE-conjugated Mouse IgG1 isotype control.

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user

Clone Ki-67 is a mouse monoclonal antibody that detects proliferation marker protein Ki-67.

Lyophilized from PBS containing D-Mannitol and Sucrose, normal appearance is a dried pellet. Reconstituted antibody solution is stable and functional as assessed by functional testing. Contains no biocide or preservatives, such as azide.

1.0 mg/mL after reconstitution at 25 μL in PBS. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.

Recommend storage of lyophilized product at 2-8°C. Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial. Reconstitute each vial by adding 25 μL PBS. Protect from light.

ColorWheel is a registered trademark of Merck KGaA, Darmstadt, Germany

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.